Team:BYU Provo/Notebook/Cholera - Enzyme/May-June/Period2/Dailylog

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Cholera - Enzymes Notebook: May 15 - May 31 Daily Log



Overview
March-April
May-June
July-August
September-October

5/15/13

We ran phusion PCR samples on regular agarose gel. See products results below

[IMAGE]

We need to re-setup the PCR products as there was no visible band for either the AmyA or Savinase proteins on our gel.


5/16/13

We reran phusion PCR for AmyA and Savinase. In addition, we also re-boiled the DNA template for Subtilisin savinase. In the PCR master mix we used a GC base thermobuffer in place of the normal 5X Phusion buffer. We hope this will help the PCR reaction for our enzymes.


5/17/13

We ran our phusion PCR products on agarose gel. See the results below.

[IMAGE]

Row 1: AmyA control
Row 2: AmyA sample 1
Row 3: Amy A sample 2
Row 4: Savinase control
Row 5: Savinase sample 1
Row 6: Savinase sample 2

The AmyA products worked. There was no visible band for the Savinase products. It seems like we are not getting adequate DNA from our lysed Bacillus Subtilis cells. We need to determine a new boiling method to better isolate the Savinase DNA in preparation for the PCR.




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